Microparticles from apoptotic vascular smooth muscle cells induce endothelial dysfunction, a phenomenon prevented by β3-integrin antagonists

Summary

Fragile atherosclerotic plaques are rich in apoptotic smooth muscle cells (SMCs) and macrophages, generating microparticules (MPs) which accumulate locally and may be released in blood in case of mechanical or spontaneous plaque disruption. Besides being highly procoagulant, this material may interact with downstream endothelium. Using a model of mouse aorta vaso-reactivity, we have investigated the effects of apoptotic MPs preparedin vitro from Fas-ligand sensitive SMCs. Short-term preincubation of aorta rings with the MPs dose-dependently reduced the vasodilatory response to acetylcholine dependent on the endothelium. This effect was prevented by the addition of abxicimab or eptifibatide, indicating a role for a β3 integrin in thisprocess. We further investigated its mechanism using cultured endothelial cells. The MPs were found to bind to the cells and to inhibit the production and the release of nitric oxide (NO) in response to bradykinin. This phenomenom was redox sensitive, independent of the generation of activated coagulation proteases, and was abrogated when the MPs were pretreated by trypsin. The metabolic effects of MPs were prevented by the addition of eptifibatide. Taken together, these results suggest a potential, platelet-independent, mechanism for the improvement of microvascular perfusion observed with β3-integrin antagonists.

Keywords

Microparticles - nitric oxide - endothelium - vasorelaxation - β3-integrin antagonists

^* S. E. and A. B. contributed equally to this work

• References

• 1 Rezkalla SH, Kloner RA. No-reflow phenomenon. Circulation 2002; 105: 656-62.
• 2 Bonderman D, Teml A, Jakowisch J. et al. Coronary no-reflow is caused by shedding of active tissue factor from dissected atherosclerotic plaque. Blood 2002; 99: 2794-800.
• 3 Kotani J, Nanto S, Mintz GS. et al. Plaque gruel of atheromatous coronary lesion may contribute to the noreflow phenomenon in patients with acute coronary syndrome. Circulation 2002; 106: 1672-7.
• 4 Mallat Z, Hugel B, Ohan J. et al. Shed membrane microparticles with procoagulant potential in human atherosclerotic plaques: a role for apoptosis in plaque thrombogenicity. Circulation 1999; 99: 348-53.
• 5 Brisset AC, Terrisse AD, Dupouy D. et al. Shedding of active tissue factor by aortic smooth muscle cells (SMCs) undergoing apoptosis. Thromb Haemost 2003; 90: 511-8.
• 6 Ghrib F, Brisset AC, Dupouy D. et al. The expression of tissue factor and tissue factor pathway inhibitor in aortic smooth muscle cells is up-regulated in synthetic compared to contractile phenotype. Thromb Haemost 2002; 87: 1051-6.
• 7 Mallat Z, Benamer H, Hugel B. et al. Elevated levels of shed membrane microparticles with procoagulant potential in the peripheral circulating blood of patients with acute coronary syndromes. Circulation 2000; 101: 841-3.
• 8 Boulanger CM, Scoazec A, Ebrahimian T. et al. Circulating microparticles from patients with myocardial infarction cause endothelial dysfunction. Circulation 2001; 104: 2649-52.
• 9 Morel O, Hugel B, Jesel L. et al. Sustained elevated amounts of circulating procoagulant membrane microparticles and soluble GPV after acute myocardial infarction in diabetes mellitus. Thromb Haemost 2004; 91: 345-53.
• 10 Nomura S, Uehata S, Saito S. et al. Enzyme immunoassay detection of platelet-derived microparticles and RANTES in acute coronary syndrome. Thromb Haemost 2003; 89: 506-12.
• 11 Bernal-Mizrachi L, Jy W, Jimenez JJ. et al. High levels of circulating endothelial microparticles in patients with acute coronary syndromes. Am Heart J 2003; 145: 962-70.
• 12 Barry OP, Pratico D, Lawson JA. et al. Transcellular activation of platelets and endothelial cells by bioactive lipids in platelet microparticles. J Clin Invest 1997; 99: 2118-27.
• 13 Nomura S, Tandon NN, Nakamura T. et al. Highshear-stress-induced activation of platelets and microparticles enhances expression of cell adhesion molecules in THP-1 and endothelial cells. Atherosclerosis 2001; 158: 277-87.
• 14 Martin S, Tesse A, Hugel B. et al. Shed membrane particles from T lymphocytes impair endothelial function and regulate endothelial protein expression. Circulation 2004; 109: 1653-9.
• 15 Mesri M, Altieri DC. Leukocyte microparticles stimulate endothelial cell cytokine release and tissue factor induction in a JNK1 signaling pathway. J Biol Chem 1999; 274: 23111-8.
• 16 Brodsky SV, Zhang F, Nasjletti A. et al. Endothelium-derived microparticles impair endothelial function in vitro. Am J Physiol Heart Circ Physiol 2004; 286: H1910-5.
• 17 Chiu JJ, Chen LJ, Lee PL. et al. Shear stress inhibits adhesion molecule expression in vascular endothelial cells induced by coculture with smooth muscle cells. Blood 2003; 101: 2667-74.
• 18 Huber J, Vales A, Mitulovic G. et al. Oxidized membrane vesicles and blebs from apoptotic cells contain biologically active oxidized phospholipids that induce monocyte-endothelial interactions. Arterioscler Thromb Vasc Biol 2002; 22: 101-7.
• 19 Shaul PW. Endothelial nitric oxide synthase, caveolae and the development of atherosclerosis. J Physiol 2003; 547: 21-33.
• 20 Doi H, Kugiyama K, Ohgushi M. et al. Remnants of chylomicron and very low density lipoprotein impair endothelium-dependent vasorelaxation. Atherosclerosis 1998; 137: 341-9.
• 21 Doi H, Kugiyama K, Oka H. et al. Remnant lipoproteins induce proatherothrombogenic molecules in endothelial cells through a redox-sensitive mechanism. Circulation 2000; 102: 670-6.
• 22 Moiseeva EP. Adhesion receptors of vascular smooth muscle cells and their functions. Cardiovasc Res 2001; 52: 372-86.
• 23 Lele M, Mansoor S, Wajih N. et al. Eptifibatide and 7E3, but not tirofiban, inhibit ανβ3 integrin mediated binding of smooth muscle cells to thrombospondin and prothrombin. Circulation 2001; 104: 582-7.
• 24 Dorfleutner A, Hintermann E, Tarui T. et al. Crosstalk of integrin alpha3beta1 and tissue factor in cell migration. Mol Biol Cell 2004; 15: 4416-25.
• 25 Silvestre JS, Thery C, Hamard G. et al. Lactadherin promotes VEGF-dependent neovascularization. Nat Med 2005; 11: 499-506.
• 26 Aymong ED, Curtis MJ, Youssef M. et al. Abciximab attenuates coronary microvascular endothelial disfunction after coronary stenting. Circulation 2002; 105: 2981-5.
• 27 Steiner S, Seidinger D, Huber K. et al. Effect of glycoprotein IIb/IIIa antagonist abciximab on monocyteplatelet aggregates and tissue factor expression. Arterioscler Thromb Vasc Biol 2003; 23: 1697-702.
• 28 Heitzer T, Ollmann I, Köke K. et al. Platelet glycoprotein IIb/IIIa receptor blockade improves vascular nitric oxide bioavailability in patients with coronary artery disease. Circulation 2003; 108: 536-41.